Method for treating cholesterolemia by administering resorcylic acid lactone derivatives

ABSTRACT

A process, and the composition used therein, for estrogenic therapy which comprises administering daily to the patient from about 0.2 to 2000 milligrams of a compound having the formula: ##STR1## wherein R is selected from the group consisting of hydrogen, lower alkyl and a lower saturated acyclic acyl; Z is selected from the group consisting of &gt;CH 2 , &gt;CHOH and &gt;C=O; and A is selected from the group consisting of --CH 2  --CH 2  -- and --CH=CH--.

This is a division of application Ser. No. 576,639, filed May 12, 1975,now U.S. Pat. No. 3,965,275, which in turn is a continuation ofapplication Ser. No. 441,150, filed Feb. 11, 1974, abandoned; which inturn is a continuation of application Ser. No. 289,456, filed Sept. 15,1972, abandoned; which in turn is a continuation of application Ser. No.28,913, filed Apr. 15, 1970, abandoned; which in turn is acontinuation-in-part of application Ser. No. 512,199, filed Dec. 7,1965, abandoned.

The present invention is directed to a pharmaceutical composition usefulin carrying out estrogenic therapy for human patients.

The administration of estrogenic hormones such as estradiol andestrogenic substances such as diethylstilbestrol in the therapeutictreatment of various illnesses and disorders such as acne, capillaryfragility, emotional instability, delayed puberty, reduction of bloodcholesterol and habitual abortion, has been practiced for years withconsiderable success. The estrogen hormones and estrogenic substancescurrently available are not without criticism, however. A major drawbackof these substances, particularly as applied to their administration inmen, is their strong "feminizing" effect. Many women also manifestundesirable side-effects from these compounds, as, for example,excessive menstrual flow and masculinization. A search has been inprogress for years with little success for a "weak estrogen" compoundwhich is effective in estrogenic therapy but lacks distinct feminizingand other undesirable side effects.

In accordance with the present invention, there is provided apharmaceutical composition containing an effective therapeuticallyactive estrogenic ingredient characterized by relatively weak feminizingand other side effect estrogenic activity.

The active ingredient of the pharmaceutical compositions can berepresented by the structural formula: ##STR2## wherein R is hydrogen,substituted or unsubstituted alkyl, e.g. lower alkyl such as methyl,ethyl, hexyl, etc., and acyl, e.g., lower saturated acyclic acylradicals such as acetyl and valeryl; A is --CH₂ --CH₂₋₋ or --CH=CH-- andZ is >C=O, >CH₂ or >CHOH. As will be demonstrated below, the compoundsof the invention when compared with diethylstilbestrol, a compound ofknown estrogenic activity used in estrogenic therapy in the mouseuterotropic activity test, exhibit effective but relatively weakestrogenic activity.

The compounds of the present invention include the compound: ##STR3##hereinafter referred to as the fermentation estrogenic substance(F.E.S.), from which the other compounds of the invention can beproduced by reduction of the ketone group to replace the oxygen of theketone group with two hydrogen atoms, by reducing the ketonic group toadd two hydrogens thereto, by saturation of the olefinic bond or anycombination of such reductions. The reduction of the ketone group toreplace the oxygen can be effected by several procedures. One of theseprocedures involves the Clemmensen reduction using zinc and hydrochloricacid; another involves the Wolff-Kishner reduction using hydrazine andalkali, e.g. NaOH, and the third involves formation of the dithioacetalwith ethylene dithiol or ethylmercaptan and the catalyticdesulfurization with Raney nickel catalyst containing adsorbed hydrogen.

The addition of two hydrogen atoms to the ketonic group and saturationof the olefinic bond can be obtained by conventional reductionprocedures, for instance, in the presence of Raney nickel catalyst. Thereduction is preferably carried out with the F.E.S. suspended ordissolved in a suitable solvent, e.g. an alcohol, preferably a loweralkanol such as methanol, ethanol, etc. In general, the reduction can beaccomplished at ambient temperatures and ambient pressures. Preferabletemperatures are from about 15° to 40° C., and preferable pressures areof from about 1 to 100 atmospheres. In general, from about 0.1 to 5grams of catalyst are used per gram of F.E.S.

In producing compounds of the invention where A is --CH₂ --CH₂ -- theolefinic bond of F.E.S. can be reduced, for example, by hydrogenation inthe presence of a Group VIII metal, particularly platinum or palladiumcatalyst on a suitable carrier, e.g., charcoal. Generally, the catalystcontains from about 0.01 to about 10% of the catalytic metal. Thecatalyst is used in a ratio of generally from 0.02 to 2 grams ofcatalyst, preferably about 0.1 to 0.5 gram, and particularly about 0.2gram catalyst per gram of F.E.S. The reduction may be carried out whilethe F.E.S. is dissolved in a suitable solvent, e.g. an alcohol,especially a lower alkanol such as 2-propanol, methanol, ethanol, andacid, e.g. acetic acid, etc. at ambient temperatures; e.g. from about15° to 40° C., and ambient pressures, since only the presence ofhydrogen is required; however, it is preferred to utilize an elevatedpressure, e.g. from about 1 to 50 atmospheres of hydrogen.

In producing compounds of the present invention where R is alkyl,conventional alkylation procedures can be used to replace the H atom ofone or both of the OH groups on the benzene ring of F.E.S. with an alkylgroup. Alkylated dihydro F.E.S. compounds can be produced, for example,by first alkylating F.E.S. and then reducing it as set forth supra, orby first reducing it and then alkylating it. The alkylation can be byreaction with the corresponding dialkyl sulfates, e.g. dimethyl sulfate,diethyl sulfate, etc. to produce the dialkyl F.E.S. or a monoalkylF.E.S. with the alkyl group replacing the hydrogen of the hydroxyl groupon the benzene ring ortho to the ester group. Furthermore, a monomethylF.E.S. compound with the methyl group replacing the hydrogen of thehydroxyl group para to the ester group can be selectively produced usingdiazomethane.

In producing compounds of the present invention where R is acyl,conventional acylation procedures can be used to replace the hydrogenatom of one or both of the hydroxyl radicals on the benzene ring ofF.E.S. with an acyl radical. Acylated F.E.S. compounds can be produced,for example, by reaction with the corresponding acid anhydride, e.g.acetic anhydride, propionic anhydride, etc., catalyzed with, forexample, sodium acetate or pyridine. Ambient conditions can be usedalthough it is preferred to keep the reaction mixture cold. Whencompounds having one R as alkyl and the other acyl, it is advantageousto alkylate before acylating.

The fermentation estrogenic substance (F.E.S.) is so named since aconvenient method for producing it is by cultivating, on a suitablenutrient medium, the organism Gibberella zeae (Gordon) on deposit at theNorthern Utilization Research and Development Division of the UnitedStates Department of Agriculture under the number NRRL-2830.

Specific examples of the preparation of F.E.S. and other compounds ofthe invention are given below and disclosed in more detail in U.S. Pat.Nos. 3,196,019; 3,239,354; 3,239,345 and 3,239,341.

The pharmaceutical compositions of the invention can be prepared bymixing the active ingredient with non-toxic, pharmaceutically-acceptablecarriers, which can be inert diluents or solid carriers, and forming theresulting mixture into suitable dosage unit forms. The compositions canbe administered to the subject by any suitable method including oral andparenteral administration. Forms suitable for oral administrationinclude, for example, pressed or coated tablets, capsules or pills,syrups, solutions or suspensions in water or non-toxic organic solventmedia such as propylene glycol and glycerol formal, and dispersiblepowders. Compositions suitable for parenteral administration are theknown pharmaceutical forms for such administrations, for example,sterile aqueous suspensions or solutions in oil media. The sterileaqueous suspensions can be formulated in the presence of parenterallyacceptable buffers, e.g. sodium citrate, citric acid and/orpreservatives such as phenol and methyl and propyl esters of p-hydroxybenzoic acid. A preferred oily media for preparation of the sterileaqueous solution is peanut oil. For treatment of the skin, the activeingredient can be mixed in any of the conventional cosmetic base lotionsin a hydrophilic base.

The pharmaceutical compositions may also include adjuvants known in theart as desirable or useful as, for example, wetting agents, dispersingagents, suspending agents, lubricating agents, sweetening agents,coloring agents and flavoring agents.

Illustrative or oral compositions are tablets wherein the activeingredient is mixed with inert fillers, e.g., dicalcium phosphate, terraalba or lactose in the presence of disintegrating agents, as, forexample, maize starch and in the presence of lubricating agents such asmagnesium stearate. Examples of suitable aqueous solutions for oral useare those formulated by incorporating the active ingredient in inertpharmaceutically-acceptable liquid solvent media which can contain, ifdesired, pharmaceutically-acceptable thickening agents such as sodiumcarboxymethyl-cellulose and/or pharmaceutically-acceptable sweeteningand flavoring agents.

The actual amounts of the active ingredient in the pharmaceuticalcomposition of the invention may vary depending on the particulardisorder treated but in all cases the amount present is that sufficientto produce the desired therapeutic effect. In general, for carrying outestrogenic therapy for a human patient from about 0.2 to 2000 milligramsof the active ingredient are administered daily. This amount isadministered in amounts of from about 0.2 to 500 mg. preferably 1.0 to100 mg. per dosage unit. More particularly, in post-menopause usage theactive ingredient will be administered in a range of about 0.1 to 10mg., preferably about 0.5 to 6 mg. per day per kilogram body weight ofthe patient. With females this dosage is typically administered in amanner similar to oral contraceptives, that is, 20 to 25 days on thedrug followed by rest periods of up to 10 days, e.g. 5 to 10 days. Oftenthis cycle is referred to as 3 weeks on the drug and 1 week rest period.Post-menopause usage and the dosages used therefore includes treatmentof capillary fragility and emotional instability and such dosages arethose used with habitual abortion, delayed puberty, etc. The cycleregime is used to avoid building of endometrium as is well known. Thedosage for treatment of cholesterol, i.e. for cholesterol lowering, isfrom about 0.5 to 20 mg./day/kilogram of body weight and the dosage isusually given continuously although cyclic administration can be used.Acne is treated with a skin cream containing the active ingredient, e.g.0.1 to 10 wt. % of the active ingredient in a cosmetic base lotion or ina hydrophilic base.

The following examples are offered to illustrate this invention;however, the invention is not limited to the specific materials, amountsand procedures set forth. The first example illustrates preparation of asuitable inoculum containing the organism Gibberella zeae (Gordon)NRRL-2830.

EXAMPLE 1

A spore sand culture containing Gibberella zeae (Gordon) NRRL-2830 wasaseptically placed in a sterile tube containing 15 milliliters ofCzapek's-Dox solution and a small amount of agar. This medium was thenincubated for about 168 hours at approximately 25° C. At the end of theincubation period, the medium was washed with 5 milliliters of steriledeionized water and transferred to a sterile tube containing 45milliliters of Czapek's-Dox solution. The contents of the tube were thenincubated for about 96 hours at about 25° C. after which the materialwas available for use in inoculation of a fermentation medium.

The following example illustrates the fermentation of the organismGibberella zeae (Gordon) NRRL-2830 to produce F.E.S.

EXAMPLE II

To a 2 liter flask was added 300 grams of finely divided corn. The flaskand its contents were then sterilized and after sterilization 150milliliters of sterile deionized water were added. To the mixture in theflask were then added 45 milliliters of the inoculum prepared by theprocess of Example I and the material was thoroughly mixed. The mixedmaterial was then incubated for about 20 days at 25° C. in a dark roomin a water-saturated atmosphere.

The following example illustrates the recovery of the F.E.S. from thefermentation medium.

EXAMPLE III

A 300 gram portion of fermented material produced by the method ofExample II was placed in 500 milliliters of deionized water andslurried. The slurry was then heated for about 15 minutes at 75° C., 300grams of filter aid were then added and the material was filtered. Thesolid filtered material containing the F.E.S. was then air dried, and333 grams of the dried cake were then extracted with 500 milliliters ofethanol. This procedure was repeated three more times. The ethanolextract was evaporated to dryness under vacuum to give 6.84 grams ofsolid material. This solid material was then dissolved in 20 millilitersof chloroform and extracted with 30 milliliters of an aqueous solutioncontaining 5% by weight of sodium carbonate having an adjusted pH ofabout 11.2. The extraction process was repeated seven more times. The pHof the sodium-carbonate extract was then adjusted to 6.2 withhydrochloric acid, to yield a F.E.S. substance-containing precipitate.The precipitate and the aqueous sodium-carbonate extract were then eachin turn extracted with 75 milliliters of ethyl ether. This procedure wasrepeated three more times to yield a light yellow ethereal solution,which was then evaporated to yield 116 milligrams of solid F.E.S. Thismaterial was then subjected to multiple transfer countercurrentdistribution using 100 tubes and a solvent system consisting of twoparts chloroform and two parts carbon tetrachloride as the lower phaseand four parts methanol and one part water as the upper phase, all partsby volume. The solid material obtained from the multiple transfercountercurrent distribution was F.E.S.

The following examples, Examples IV to VI, illustrate the reduction ofF.E.S. to produce tetrahydro F.E.S. having the formula: ##STR4##

EXAMPLE IV

Tetrahydro F.E.S. was produced by dissolving 0.5 gram F.E.S. in 200milliliters of ethanol. The F.E.S. was reduced by contacting thesolution with hydrogen for 3 hours at 30° C. with 1000 psi using 2 gramsof Raney nickel as a catalyst. After filtering and concentrating thereaction mixture, the product was washed with 2 to 3 milliliters of2-nitropropane and crystallized. It was found to have a melting pointfrom 143° -160° C.

EXAMPLE V

The reduction of F.E.S. was conducted in methanol at 30° C. and 1000 psihydrogen pressure for 5 hours using Rnaey nickel catalyst to provide aproduct melting, after several crystallizations from 2-nitropropane andnitromethane, at 141°-143° C. and analyzing:

    ______________________________________                                        Calc.       (C.sub.18 H.sub.26 O.sub.5)                                                                  Found                                              ______________________________________                                        % C         67.1          67.2                                                % H         8.14          8.28                                                ______________________________________                                    

EXAMPLE VI

The reduction of 1 gram of F.E.S. was conducted in 150 cc. of ethanol atroom temperature and 50 psi of hydrogen for 4 hours in the presence of asmall amount of Raney nickel (about 1 cc. of a thick suspension inwater). The product was concentrated, treated with 5 milliliters ofisopropyl alcohol, cooled and filtered. The filtrate was mixed with 5milliliters of water, left standing over night, cooled and filtered toprovide 0.65 gram of product having a melting point of 147°-157° C. Thisproduct was recrystalized from isopropyl alcohol-water mixtures twotimes to provide 0.18 gram of a product having a melting point of178°-180° C. A product having a melting point of 146°-148° C. andweighing 0.22 gram was also recovered from the filtrate after the firstrecrystallization of the product weighing 0.65 gram. The reduction ofthe ketone group introduces an asymmetric carbon atom and makesdiastereoisomers possible. The optical activities of the two productswere (1) for THFES(HM), the product with a melting point of 178°-180°C., [α]_(D) ²⁵ = about + 46° e.g. and (2) for a combination of theTHFES(HM) and THFES(LM), the combination product having a melting pointof 146°-148° C., [α]_(D) ²⁵ = about +39° e.g. where [α] = (α .100/c.1),c = 1% in methanol and 1=2 dcm. This product is actually a mixture(about 1:2) of the product melting at 178°180° C. and its isomer. Thelow melting isomer can be obtained in pure farm by recrystallization outof the combination product using glacial acetic acid to provide pureTHFES(LM) which melts at ˜ 155° C. and has optical activity [α]_(D) ²⁵=+36°.

The following example illustrates the preparation of deoxy tetrahydroF.E.S.

EXAMPLE VII

Two 10 gram portions of F.E.S., each in 200 milliliters acetic acid,were catalytically reduced at room temperature in the presence of 1.2grams of PdO catalyst at a hydrogen pressure of about 45 psi. Thecombined reduction mixtures were heated to boiling, filtered, and thefilter cake was washed with 50 milliliters of hot acetic acid. Thecooled filtrate was added, with stirring, to 2 liters of water. Themixture was stirred for 15 minutes and the white solid was collected byfiltration, washed and dried in a vacuum desiccator to yield 19.1 gramsof dihydro F.E.S. in which the ethylenic unsaturation is saturated, andhaving a melting point of 191°-193° C.

The dihydro F.E.S. (1 gram) is added slowly with cooling (ice-bath), toa mixture of 5 cc. of ethylene dithiol 0.025 gram of freshly fused zincchloride and 2 grams of anhydrous sodium sulfate, contained in amicroflask. The mixture is maintained at 5° C. for 20 hours and then atroom temperature for 4 hours, whereupon it is poured into 50° cc. of iceand the precipitate is collected and subjected to hydrogenolysis. To thereaction product is added 100 cc. of 90% ethanol and 15 grams of Raneynickel catalyst and the mixture is refluxed until the reaction iscomplete. The nickel is removed by centrifugation and is washed severaltimes with hot ethanol by centrifugation followed by decantation, andthe centrifugates are combined. The mixture is evaporated to dryness andthe residue is suitably recrystallized to yield deoxy THFES having theformula: ##STR5##

EXAMPLE VIII

Nitrosomethylurea in an amount of 1.2 grams was slowly added to a coldmixture of 3.6 milliliters of 50% potassium hydroxide and 17 millilitersof ether. After a few minutes the yellow ether layer of the mixture wasdecanted, dried over potassium hydroxide, and then added to a solutionof 0.30 gram F.E.S. in 17 milliliters of ether. The resulting yellowmixture was left overnight in a loosely stoppered flask and then etherand diazomethane were evaporated using a steam bath. The remaining gummyresidue was crystallized by adding 3 milliliters of water, heating to60° C., and adding ethanol almost to solution. On cooling, crystalsformed yielding 0.137 gram of a product having a melting point of111°-122° C. and analyzing:

    ______________________________________                                        Calc.       (C.sub.19 H.sub.24 O.sub.5)                                                                 Found                                               ______________________________________                                        % C         68.7          68.3                                                % H         7.28          7.38                                                % OCH.sub.3 9.34          9.17                                                ______________________________________                                    

The p methyl F.E.S. is substituted for the F.E.S. in followingessentially the same procedure used in Example VII to produce a compoundhaving the formula: ##STR6##

The following example illustrates the production of dimethyl F.E.S. andmonomethyl F.E.S. derivatives, the monomethyl F.E.S. derivative havingthe hydrogen in the hydroxyl group ortho to the ester group replacedwith a methyl group.

EXAMPLE IX

Dimethyl sulfate (5 milliliters) was added to a solution of 2.24 gramsof F.E.S. in 80 milliliters 10% NaOH and 20 milliliters water. Themixture was stirred for one-half hour at 18°-20° C. (cooling bath) andan additional 5 milliliters of dimethyl sulfate was added. After anadditional 70 minutes of stirring at 20°-26° C., the solid precipitate,Solid A, was collected by filtration, washed with water and dried in avacuum desiccator. The filtrate from Solid A was acidified with 25milliliters 12 N H₂ SO₄ to yield a second precipitate, Solid B, whichwas collected, washed with water, and dried.

Solid A (0.79 gram having a melting point of 114°-118° C.) wasrecrystallized from a mixture of 10 milliliters water and 15 millilitersethanol to yield 0.66 gram of dimethyl F.E.S. having a melting point of108°-110° C.

Solid B (1.39 grams having a melting point of 152°-162° C.) wasrecrystallized twice from a mixture of water and alcohol to yield 0.8gram of monomethyl F.E.S. having a melting point of 169°-174° C. and thefollowing analysis of recrystallized Solid B (monomethyl F.E.S.) wasobtained:

    ______________________________________                                                 Calc. (C.sub.19 H.sub.24 O.sub.5)                                                            Found                                                 ______________________________________                                        % C        68.65           67.97                                              % H        7.28            7.16                                               % OMe      9.34            9.28                                               ______________________________________                                    

Each of the o methyl F.E.S. and the dimethyl F.E.S. is substituted forthe F.E.S. in the procedure of Example VII to produce the respectivecompounds: ##STR7##

The following example illustrates the production of an acylatedmonomethyl F.E.S. derivative.

EXAMPLE X

To a solution of 368 milligrams of p methyl F.E.S. in 8 milliliterspyridine is added 5 milliliters acetic anhydride and the mixture is heldat room temperature for 16 hours. Twenty-five milliliters of water arethen added. The mixture is stored in a refrigerator for 2 hours. Thesolid precipitated is collected by filtration, washed with water anddried in a vacuum desiccator to recover a compound which is substitutedfor the F.E.S. in the procedure of Example VII to produce a compound ofthe formula: ##STR8## which is recovered.

EXAMPLE XI

The compound: ##STR9## is produced by substituting o methyl F.E.S. forthe dihydro F.E.S. in the procedure of Example VII.

The following example illustrates the reduction of F.E.S. to producedihydro F.E.S. having the structure: ##STR10##

EXAMPLE XII

Two 10-gram portions of F.E.S. each in 200 milliliters acetic acid werecatalytically reduced at room temperature in the presence of 1.2 gramsof PdO catalyst at a hydrogen pressure of about 45 psi. The combinedreduction mixtures were heated to boiling, filtered, and the filter cakewas washed with 50 milliliters of hot acetic acid. The cooled filtratewas added, with stirring, to 2 liters of water. The mixture was stirredfor 15 minutes and the white solid was collected by filtration, washedand dried in a vacuum desiccator to yield 19.1 grams of dihydro F.E.S.having a melting point of 191°-193° C.

The production of dimethyl dihydro F.E.S. is illustrated by thefollowing Example.

EXAMPLE XIII

Dihydro F.E.S. (556 milligrams) was dissolved in 25 milliliters 10% NaOHand 10 milliliters water and the solution was stirred. To the stirredsolution was added 3, 2-milliliter portions of dimethyl sulfate athalf-hour intervals followed by stirring for an additional hour. Themixture was acidic and it was made alkaline by the addition of 10milliliters 10% NaOH and the alkaline mixture was stirred one-half hour.The solid formed was collected by filtration, washed with water anddried in a vacuum desiccator. The product weighed 526 milligrams andmelted at 115°-117° c. Recrystallization from a mixture of 10milliliters of water and 25 milliliters of ethanol provided 371milligrams of material having a melting point of 124°-125.5° C. It wasanalyzed with the following results:

    ______________________________________                                                 Calc. (C.sub.20 H.sub.28 O.sub.5)                                                            Found                                                 ______________________________________                                        % C        68.95           69.02                                              % H        8.10            8.12                                               % CH.sub.3 O                                                                             17.81           17.81                                              ______________________________________                                    

The following example illustrates the production of monomethyl anddimethyl dihydro F.E.S., the monomethyl dihydro F.E.S. having a methylgroup which replaced the hydrogen of the hydroxyl group on the benzenering ortho to the ester group.

EXAMPLE XIV

Dimethyl sulfate (5 ml.) was added to a solution of 2.24 g. F.E.S. in 80ml. of a 10% NaOH solution and 20 ml. of water. The mixture was stirredfor one-half hour at 18°-20° C. (cooling bath) and an additional 5 ml.of dimethyl sulfate was added. After an additional 70 minutes ofstirring at 20°-26° C., the solid precipitate, Solid A, was collected byfiltration, washed with water and dried. The filtrate from Solid A wasacidified with 25 ml. 12N H₂ SO₄ to yield a second precipitate, Solid B,which was collected, washed with water, and dried.

Solid A (0.79 g. having a melting point of 114°-118° C.) wasrecrystallized from a mixture of 10 ml. water and 15 ml. ethanol toyield 0.66 g. of dimethyl F.E.S. having a melting point of 108°-110° C.

Solid B (1.39 g. having a melting point of 152°-162° C.) wasrecrystallized twice from a mixture of water and alcohol to yield 0.80g. of monomethyl F.E.S. product having a melting point of 169°-174° C.Analysis of Solid B showed:

    ______________________________________                                                 Calc. (C.sub.19 H.sub.24 O.sub.5)                                                            Found                                                 ______________________________________                                        % C        68.65           67.97                                              % H        7.28            7.16                                               % OMe      9.34            9.28                                               ______________________________________                                    

The olefinic bond of each of the dimethyl F.E.S. and monomethyl F.E.S.is reduced using 50 psi of hydrogen and a small amount of 5% Pd oncharcoal catalyst in ethanol and conducting the reduction for 3 hours.

EXAMPLE XV

Monomethyl F.E.S. with the methyl group replacing the hydrogen of thehydroxyl group on the benzene ring para to the ester group was preparedby the following procedure:

Nitrosomethylurea in an amount of 1.2 grams was slowly added to a coldmixture of 3.6 milliliters of 50% potassium hydroxide and 17 millilitersof ether. After a few minutes the yellow ether layer of the mixture wasdecanted, dried over potassium hydroxide, and then added to a solutionof 0.30 grams F.E.S. in 17 milliliters of ether. The resulting yellowmixture was left overnight in a loosely stoppered flask and then etherand diazomethane were evaporated off using a steam bath. The remaininggummy residue was crystallized by adding 3 milliliters of water, heatingto 60° C., and adding ethanol almost to solution. On cooling, crystalsformed, yielding 0.137 grams of a product having a melting point of111°-116° C. which was again recrystallized in the same way to yield0.082 grams of monomethyl F.E.S. having a melting point of 120°-122° C.and the following analysis:

    ______________________________________                                        Calc.       (C.sub.19 H.sub.24 O.sub.5)                                                                 Found                                               ______________________________________                                        % C         68.7          68.3                                                % H         7.28          7.38                                                % OCH.sub.3 9.34          9.17                                                ______________________________________                                    

The olefinic bond of this compound is reduced according to the procedureof Example IX.

The following example demonstrates the relatively low estrogenicactivity of the compounds of the pharmaceutical compositions of theinvention.

EXAMPLE XVI

A solution of each test compound identified in the table below wasadmixed with a standard pulverized mouse ration and the solvent wasremoved by evaporation to provide a dry ration containing the level oftest compound per gram of feed indicated in the table below. A controlration and each test ration were fed to 5 to 10 ovariectomized miceweighing about 20 to 23 grams each at a level of 3 grams per day for aperiod of 5 days after which the mice were sacrificed and their uteriweighted. An increase in the weight of the uteri from the animal fed thetest compounds over the weight of the uteri in the control animalsdemonstrates estrogenic activity for the test compound.

The control animals showed a percent uterine weight to body weight ofabout 0.048 to 0.050. A uterine response of 0.060 being justsignificant, the dose required to give a response of 0.060 then givesthe relative uterotrophic activity. The following results were obtainedfrom curves drawn plotting dose/uterus weight as percent body weight.

    __________________________________________________________________________                           Uterotrophic Activity Relative To                      __________________________________________________________________________              Estimated                                                           Compound  M.E.D.*      DES       FES                                          __________________________________________________________________________    DES+      0.003 μg/g feed                                                                         1         2000                                         HMTHFES.sup.(1)                                                                         1.25         .0024     4.8                                          LMTHFES.sup.(2)                                                                         2.25         .0013     2.7                                          DeoxyTHFES.sup.(3)                                                                      6.25         .00048    0.96                                         Dimethyl FES.sup.(4)                                                                    100.0 (estimated)                                                                          .000003   0.06                                         4-methyl FES.sup.(5)                                                                    100.0 (estimated)                                                                          .000003   0.06                                         2-methyl FES.sup.(6)                                                                    28.0         .000017   0.22                                         Dihydro FES.sup.(7)                                                                     3.0          .0010     2.0                                          FES       6.0          .0005     1.0                                          __________________________________________________________________________     +diethylstilbestrol                                                           *minimal effective dosage                                                     .sup.(1) high melting tetra hydro FES prepared as in Example VI               .sup.(2) low melting tetra hydro FES prepared as in Example V                 .sup.(3) prepared as in Example VII                                           .sup.(4) prepared as in Example IX                                            .sup.(5) prepared as in Example VIII                                          .sup.(6) prepared as in Example IX                                            .sup.(7) prepared as in Example XII                                      

EXAMPLE XVII

Pharmaceutical preparation containing the compound of Example IV in theform of tablets suitable for administration to human patients;

246 grams of THFES(HM) is triturated with 60 grams of lactose to form anhomogeneous powder. To the powder is added 20 grams of silicic acid withhydrolyzed starch and water and the mixture stirred until a homogeneouspaste is formed. The paste is then dried and tabletted with 2 gramsmagnesium stearate to form tablets containing approximately 150 mg. ofactive ingredient. Similar compositions including F.E.S., THFES(LM),deoxy THFES and the alkylated and acylated compounds can be prepared bysubstitution of these compounds for THFES(HM) as the active ingredient.

EXAMPLE XVIII

Pharmaceutical preparation of aqueous suspension for oraladministration:

    ______________________________________                                        Recipe for 1000 ml. of suspension                                             ______________________________________                                        Compound of Example III 30.0      g.                                          Sucrose                 400.0     g.                                          Powdered tragacanth     7.5       g.                                          Flavoring essential oil 0.2       ml.                                         Methyl p-hydroxybenzoate                                                                              2.0       g.                                          Propyl p-hydroxybenzoate                                                                              0.5       g. .-Glycerol 150.0 ml.                     Citric acid             2.0       g.                                          Benzoic acid            1.0       g.                                          Distilled water (to complete 1000 ml.)                                        ______________________________________                                    

The glycerol, benzoic acid, methyl and propyl benzoic acids, tragacanthgum, flavoring oil and active ingredient are mixed into a homogeneousmass. An aqueous solution of the citric acid is then added with slurringand finally the sucrose is added. Slurring is continued until anhomogeneous suspension is obtained to which is added the balance of thewater. Similar compositions including F.E.S., THFES(LM), deoxy THFES andthe alkylated and acylated compounds can be prepared by substitution ofthese compounds for THFES(HM) as the active ingredient.

EXAMPLE XIX

100 mg. of the compound of Example III, 0.2 mg. ofmethyl-p-methoxybenzoate, 0.5 mg. of sodium citrate and 0.2 mg. ofcitric acid are added to 1 ml. of water. The pH of the suspension isadjusted to 5 with HCl. Heat sterilization results in an aqueoussuspension suitable for parenteral injection.

EXAMPLES XX -- XXIII

Suitable pharmaceutical compositions can be prepared by replacing theF.E.S. in the preparations of Examples XVII--XIX with any one of thecompounds of Examples IV-XV.

The following examples demonstrate the cholesterol lowering effect ofthe active ingredients of this invention.

EXAMPLE XXIV

Male Wistar rats (Royal Hart) were allowed to eat powdered PurinaLaboratory Chow ad-lib for 7 days, in order to become accustomed toeating powdered food. At a body weight of 180- 200 g, the animals werethen randomly assigned to groups of 8-10 animals. The animals werehoused in pairs in all metal cages. Throughout the "Experimental Period"of 6 days, the animals were fed ad-lib one of the following powdereddiets: (a) Plain Purina Laboratory Chow -- (controls); or (b) PurinaChow containing THFES(HM). After the 6 days experimental period, theanimals were exsanguinated by severing the external jugular veins underhexabarbital sodium acetate anesthesia (100 mg/kg I.P.) the blood beingcollected in acid-washed tubes to prevent alkaline hydrolysis of thetriglycerides. The serum was separated by centrifugation, and the totalcholesterol levels were estimated on the Auto-Analyzer using StandardMethod No. 24a for cholesterol (Technicon Auto Analyses ProceduresManual). Body weight increases were computed from measurements of bodyweight at the beginning and at the end of the experimental period.Livers were dissected and weighed wet. Absolute liver weight and liverweight as a percentage of final body weight were recorded. Foodconsumption was monitored throughout the experimental period. The actualamount of drug ingested (in mg/kg/day) was computed from the foodconsumption, final body weight, and the known amount of drug in diet.Table I below shows the effects of THFES(HM) on the serum totalcholesterol levels of the animals. Significant depression of serum totalcholesterol was seen at doses of 6.9 mg/kg/day and above. THFES(HM) didnot produce hepatomegaly under these conditions. F.E.S., THFES(LM),deoxy F.E.S. and p-methyl F.E.S. can be substituted for the THFES(HM) totreat cholesterolemia in the manner shown.

    ______________________________________                                                      Total Serum Cholesterol                                         ______________________________________                                                            Mean            Change as %                               Treatment                                                                              Dose*      ±SEM  (mg.%) of Control                                Control**                                                                              (Mg/kg/day)                                                                              58.8± 1.7    --                                        ______________________________________                                        THFES (HM)                                                                             1.0        58.5 ±                                                                              3.3    + 1.7                                     "        2.1        57.7 ±                                                                              2.4    0                                         "        6.9        49.5 ±                                                                              2.1    -13.8                                     "        8.5        46.6 ±                                                                              2.6    -19.0                                     "        13.2       41.6 ±                                                                              1.4    -27.6                                     "        16.4       38.1 ±                                                                              1.7    -34.5                                     "        30.0       33.5 ±                                                                              2.1    -41.4                                     "        53.0       33.5 ±                                                                              4.4    -41.4                                     "        100.0      24.5 ±                                                                              1.8    -57.0                                     ______________________________________                                         *Calculated from dietary concentration of drug and food consumption.          **Mean for two experiments.                                              

EXAMPLE XXV

Groups of 35-day old male rats (Holtzman) weighing 120-135 gm were heldwithout treatment for 1 week on standard laboratory ration (PurinaLabena Chow). The rats were then fed, ad libitum, for a 2-week period, adiet consisting of the following:

    ______________________________________                                        Purina Labena            50.4%                                                Cornstarch Mixture*      29.0                                                 Lard                     20.0                                                 DL-Methionine            0.6                                                  *Cornstarch Mixture:                                                           Cornstarch              70%                                                   Alphacel                12                                                    Crisco                  10                                                    USP Salt Mixture XIV    7                                                     Cod Liver Oil           1                                                    ______________________________________                                    

During the next 2 weeks (4th and 5th on experimental diet) the animalsreceived test compounds that were mixed in the above on a mg/kg of dietbasis. After exposure to this diet containing drug for 2 weeks theanimals were anesthetized, bled individually to obtain serum samples todetermine total cholesterol, triglycerides and α- and β- lipoproteins.

The results are set forth in Table II. The chemical analysis of sera ofrats on the high fat diet for 4 weeks, with drugs added for the last 2weeks of the test period, disclosed that THFES(HM) produced highlysignificant decreases in total cholesterol at all doses tested andTHFES(HM) decreased the α/β lipoprotein ratio to highly significantdegrees at the four concentrations tested; the α/β ratios decreased withincreasing concentration in the diet.

                                      Table II                                    __________________________________________________________________________            Dose    Av.Food                                                                             Total            Lipo-      Testes                              mg/kg                                                                              No.                                                                              Intake                                                                              Cholesterol                                                                           Triglycerides                                                                          proteins                                                                            α/β                                                                     Wt. Body Weight*            Compound                                                                              Diet Rats                                                                             gm/day/rat                                                                            (mg/100 ml serum)                                                                            α                                                                          β                                                                           Ratio                                                                              (gm)                                                                              Init.                                                                            Start                                                                            Final             __________________________________________________________________________    Control --   10 13    98.7±4.0                                                                           321.8±46.0                                                                          18.3                                                                             27.7                                                                             0.66 3.3 126                                                                              269                                                                              342               THPES (HM)                                                                            12.5 10 14.7  72.7±4.0***                                                                        214.2±22.2*                                                                         15.8                                                                             32.5                                                                             0.49**                                                                             3.3 127                                                                              260                                                                              287               THFES(HM)                                                                             25.0 10 13.5  62.5±3.0***                                                                        258.1±41.1                                                                          11.6                                                                             29.7                                                                             0.39**                                                                             3.2 128                                                                              264                                                                              278               "       50.0 10 12.4  59.5±5.4***                                                                        279.2±40.2                                                                          7.2                                                                              32.8                                                                             0.22***                                                                            3.1 127                                                                              255                                                                              255               "       100.0                                                                              10 11.0  52.0±2.8***                                                                        185.4±29.0                                                                          3.0                                                                              33.7                                                                             0.09***                                                                            3.1 128                                                                              260                                                                              246               __________________________________________________________________________      *P < 0.05                                                                    **P < 0.01                                                                    ***P < 0.001                                                                  *Initial = Weight on day animals were received?                                Start = Weight on day animals were placed on diet plus compound.        

While THFES(HM) produced marked decreases in the cholesterol,triglycerides and α/β- lipoprotein ratios, treatment with the compoundalso produced decreases in the weight gain of the animals. Based on thedaily food intake values, the estimated daily dose of THFES(HM) in theseanimals varied from 180 to 1100 g per rat. In addition to the findingsdiscussed above, four of 10 animals on the 12.5 mg dose level showedsigns of alopecia; 1/10 of the high dose group presented this symptom.As has been indicated in other studies there was involution of thethymus in treated animals. F.E.S. and deoxy THFES can be substituted forthe THFES(HM) to treat hypercholesterolemia in a similar manner.

The following examples demonstrate that the active ingredient of thisinvention is effective in estrogenic replacement therapy inpost-menapausal and similar treatments.

EXAMPLE XXVI

The effect of THFES(HM) on the vaginal and uterine mucosa and sex skinchanges of ovariectomized Rhesus monkeys was determined to demonstratethe post-menapausal effects of the drug. Monkeys were studied for aperiod of several days prior to drug treatment to establish base-linevaginal values. THFES(HM) was administered orally for 10 days.Observations on vaginal smear changes, coloration of the sex skin andwithdrawal bleeding were recorded during treatment and for 20 daysfollowing its cessation.

Administration of the drug resulted in increased numbers of cornifiedcells in the vaginal washings. By the fifth day of treatment with 1.8mg/kg, leukocytes were absent from the smear and remained absent for theduration of the treatment period. Coloration of the sex skin at thisdose reached its maximum on the 4th day of treatment in animal No. 13and the 11th day in animals No. 12 and No. 14. It would appear that, atthis dose level, vaginal cornification is the most sensitive indicatorof the estrogenic effect of the drug. The effect on the uterus isindicated by the withdrawal bleeding which occurred in two of threeanimals treated; there appeared to be little uniformity in the time atwhich bleeding occurred following cessation of treatment.

Four of six animals treated with 0.9 mg/kg drug had withdrawal bleedingfollowing cessation of treatment indicating uterine stimulation;"spotting" was noted in 5 of the 6 animals. This response would suggestthat 0.9 mg/kg dose was stimulating endometrial development but wasinadequate to maintain it. Vaginal changes were comparable with thoseobserved in animals treated with the 1.8 mg/kg dose level. Sex skinchanges were present and the degree of coloration was essentially thesame as was observed in the high dose group.

Animals treated with 0.45 mg/kg drug displayed minimal or noestrogen-dependent changes. Animals No. 28, No. 29 and No. 30 showed avaginal response which persisted throughout the study. Animal No. 29 had"spotting " as judged by the presence of RBC's in its smear on 11treatment or post-treatment days. Animals No. 23, No. 24 and No. 26treated at a different time, failed to respond.

Based upon the data, it can be concluded that THFES(HM) is able tostimulate estrogen-like changes in ovariectomized monkeys and can beused as a post-menapausal drug.

EXAMPLES XXVII - XXX

In each of these examples, 10 women were administered THFES(HM) atvarying dosage levels for a period of time. Tables III to VI set forththe relevant data for post-menapausal response. The administration ofthe drug and dosage was

    ______________________________________                                        TABLE       DOSAGE         TEST PERIOD                                        ______________________________________                                        Table  III      400 mg./daily  20 days                                        "      IV       200 mg./daily  20 days                                        "      V        100 mg./daily  60 days                                        "      VI        50 mg/daily   30 days                                        ______________________________________                                    

The prior symptoms appear with the patient identification and the effectof the drug is shown under Remarks. The data demonstrate theeffectiveness of THFES(HM) in relieving many of the symptoms at thedosage levels tested. At 200 mg./daily the symptoms, with the exceptionof insomnia, were relieved. At 100 mg./daily the symptoms were relievedand the patients showed excellent estrogenic response with the exceptionof Patient 63-A. There were no toxic symptoms evident. With an averageweight per patient of about 50 kg., the dosages levels are about 1 to 8mg./day/kg. body weight. The follicle stimulating hormones (FSH) data ofTable V are a clear demonstration of the activity of the drug intreatment of post-menapausal syndrome since to lower the FSH units is agood measure of such activity.

                                      TABLE III                                   __________________________________________________________________________    Vaginal Cytology                                                                                 Basal       Leuco-                                         Patient      Age                                                                              Day                                                                              Cells                                                                             CPI EI  cytes                                                                             Remarks                                    __________________________________________________________________________     39-A                                                                         Had vulvular pruritis                                                                      62  0 77% 23      Many                                                                              Shortly after start - patient              and insomnia.    7 0   38  64  Some                                                                              feeling well with slight de-               Diabetes mellitus in                                                                          14 0   56  61  v.few                                                                             crease of vulvar itching and               good control with                                                                             21 0       87  None                                                                              wetness of vulva and vagina.               diet. Biopsy-   21 Biopsy - Late prolif.                                                                         Did not have withdrawal bleed-             resting endom.  endometrium. Moderate                                                                            ing. Discontinued - lack of                                mitotic activity.  drug.                                        40-A                                                                        Suffers hot flushes                                                                        51  0 17   6  19  Some                                                                              Complete disappearance of hot              and irritability.                                                                              7 0   32  56  Some                                                                              flushes. Improvement of                    Biopsy - Early prolif,                                                                        14     45  39  v.few                                                                             psychological state. Dis-                  endom. Scarce mitotic                                                                         20     67  48  v.few                                                                             continued - lack of drug.                  activity.       20 Biopsy - Late prolif.                                                                         Withdrawal bleeding lasted                                 endometrium. Patchy                                                                              3 days.                                                    hyperplasia.                                                    41-A                                                                        Periodic diarrhea and                                                                      59  0 34   2  10      Colon condition unchanged.                 dryness of genitalia.                                                                          7 6   21  36      Genitalia moist. Has had non-              Irritability of colon                                                                         14     59  83      mal intercourse with husband.              since age 30. Biopsy-                                                                         20     48  71      Scant withdrawal bleeding                  Resting endometrium.                                                                          20 Biopsy - Late prolif.                                                                         lasting 2 days. Discontinued-                              endometrium. Good  lack of drug.                                              mitotic activity.                                               42-A                                                                        Depression, irritabil-                                                                     52 0  11  19   6  Some                                                                              Hot flushes and irritability dis-          ity and mild hot                                                                              7      24  51  Few appeared. Still depressed. No              flushes. Biopsy-                                                                              14     52  60  Few withdrawal bleeding. Discontinued          Intermed. prolif.                                                                             20     63  57  Few                                            lack of drug.                                                                 endom. Scarce   20 Biopsy - Late prolif.                                      mitotic activity.                                                                             endometrium. Good mitotic                                                     activity.                                                       43-A                                                                        Obese; has insomnia                                                                        59  0 31   7  22      No headaches. Insomnia did not             and frequent head-                                                                             7 2   32  26      improve. Slight withdrawal                 aches. Biopsy-  14     51  64      bleeding that lasted 2 days.               atrophic endom. with                                                                          20     59  47      Discontinued - lack of drug.               areas of cystic pseudo-                                                                       20 Biopsy : Late prolif.                                      hyperplasia.    endom. with moderate                                                          mitotic activity.                                               44-A                                                                        Hot flushes and dys-                                                                       51  0 24  12   8      Hot flushes disappeared. Dys-              parennia. Cholecis-                                                                            7 0   38  63      pareunia minimal. No with-                 tectomy in 1961.                                                                              14     67  100     drawal bleeding. Discon-                   Biopsy - Intermed.                                                                            20     77  91      tinued - lack of drug.                     prolif. endom.  20 Biopsy - Late prolif.                                      Scarce mitotic activity.                                                                      endometrium. Good mitotic                                                     activity.                                                       45-A                                                                                     63 0  87  marked inflammatory-                                                                      Markedly relieved - epithelia              Marked dryness of      exudate     of the genitalia are moist -               genitalia - discrete                                                                          7  14  38  22      burning sensation disappeared.             burning sensation.                                                                            14  0  63  71  Several                                                                           No withdrawal bleeding. Dis-               Has moderate hyper-                                                                           20     68  90  Some                                                                              continued - lack of drug.                  tension. Biopsy-                                                                              20 Biopsy - Late prolif.                                      atrophic endometrium.                                                                         endometrium. Moderate                                                         mitotic activity.                                               46-A       56  0 18  13  22  Several                                                                           Marked decrease of irritability;           East fatigability and                                                                          7     39  63  Some                                                                              still gets tired easily;                   irritability. Biopsy-                                                                         14     68  70  Some                                                                              Withdrawal bleeding lasted 4               resting endometrium.                                                                          20     73  69  Few days and was moderate. Dis-                                20 Biopsy - Late prolif.                                                                         continued - lack of drug.                                  endometrium. Patchy                                                           hyperplasia.                                                    47-A                                                                                     59  0 43  11   2  Some                                                                              No ill effects - had slight                No gynecological                                                                               7     43  90  Few mucorrhea lasting 2 days.                  complaints-     14     51  67  Few Withdrawal bleeding moderate               chronic colitis.                                                                              20     73  82  No  lasting 2 days. Discom-                    Biopsy - Inactive                                                                             20 Biopsy - Late prolif.                                                                         tinued - lack of drug.                     endometrium     endom. Moderate                                                               mitotic activity.                                               48-A                                                                                     52  0     23  30  Some                                                                              Abdominal discomfort and hot               Abdominal discomfort                                                                           7     53  68  v.few                                                                             flushes disappeared. Remains               rare; marked hot                                                                              14     87  82  v.few                                                                             depressed. No withdrawal                   flushes and moderate                                                                          20     79  94  v.few                                                                             bleeding. Discontinued-                    depression.     20 Biopsy - Late prolif.                                                                         lack of drug.                              Biopsy-         endometrium. Good                                             Late prolif. endom.                                                                           mitotic activity.                                             Slight mitotic ac-                                                            tivity                                                                        __________________________________________________________________________

                                      TABLE IV                                    __________________________________________________________________________    Vaginal Cytology                                                                                 Basal       Leuco-                                           Patient    Age                                                                              Day                                                                              Cells                                                                             CPI EI  cytes                                                                              Remarks                                   __________________________________________________________________________      49-A                                                                                     55  0      8   2  Few  Feels same as before starting             Discrete depression                                                                            7     28  19  Few  drug. No withdrawal bleed-                and fainting spells.                                                                          14     24  36  Few  ing. Discontinued - lack                  Biopsy - Inactive                                                                             20     31  22  Few  of drug.                                  endometrium.    20 Biopsy - intermed.                                                         prolif. endom. - Moderate                                                     mitotic activity.                                               50-A                                                                                     60  0 61          Several                                                                            Condition remained the same               Minor complaints on                                                                            7 32  21   7       during treatment, although                GF tract; bouts of                                                                            14     53  67  Some genitalia became wet.                     diarrhea. Emotionally                                                                         20     47  83  Some Discontinued - lack of drug.              unstable.       20 Biopsy - intermed.                                         Biopsy - atrophic                                                                             prolif. endom. Moderate                                       endometrium.    mitotic activity.                                                51-A                                                                                    65  0 68   2   7  Many Itching and Leukorrhea dis-               Slight itching and                                                                             7     63  70  Some appeared. Psychological                   leukorrhea; irrita-                                                                           14     56  82  Few  state did not change. Mini-               bility for past 20                                                                            20     59  67  Few  mal withdrawal bleeding for               years.          20 Biopsy - Late prolif.                                                                          one day. Discontinued-                    Biopsy - none.  endom. Moderate mitotic                                                                           lack of drug.                                             activity.                                                       52-A                                                                                     54  0 4   23  32  Some Increased wetness of vagina               Suffers dyspareunia,                                                                           7     38  40  Few  and decrease of dyspareunia.              headaches, insomnia.                                                                          14     30  31  Some Headache and insomnia not                 Biopsy - intermed.                                                                            20     47  38  Few  affected. No withdrawal                   prolif. endometrium-                                                                          20 Biopsy - Late prolif.                                                                          bleeding. Discontinued-                   few mitoses.    endometrium. Moderate                                                                             lack of drug.                                             mitotic activity.                                               53-A                                                                                     65  0 86          Many Complete disappearance of com-            Dryness of vulva                                                                               7     32  53  Many plaints. Regrets no more                  and vagina - slight                                                                           14     67  84  Some drug available. No with-                  burning sensation                                                                             20     73  69  Some drawal bleeding. Discon-                  worse at night  20 Biopsy - Late prolif.                                                                          tinued - lack of drug.                    Biopsy - atrophic                                                                             endometrium. Slight                                           endometrium     mitotic activity.                                               54-A                                                                                     53  0 22  12   4  Some No hot flushes during treat-              Marked irritability-                                                                           7  6  22  19  Some ment. First days of treat-                infrequent but very                                                                           14     48  36  Some ment moderate nausea. One                 hot flushes     20     36  49  Some episode of epigastric burn-               Biopsy - atrophic                                                                             20 Biopsy - Late prolif.                                                                          ing. Irritability did not                 endom. - cystic endometrium. Moderate                                                                             change. No withdrawal                     pseudohyperplasia.                                                                            mitotic activity.   bleeding. Discontinued-                                                       lack of drug.                               55-A                                                                                     50  0     15  23  Several                                                                            No hot flushes; palpitations              Hot flushes and  7     42  38  Few  unchanged. Epigastric                     palpitations.   14     59  64  Few  burning first few days.                   Biopsy - Intermed.                                                                            20     53  71  Few  Moderate withdrawal bleeding              prolif. endom.  20 Biopsy - Late prolif.                                                                          lasted 3 days. Discontinued-              Scaerce mitotic endometrium. Slight lack of drug.                             activity.       hyperplasia.                                                    56-A                                                                                     58  0 73          Many Marked improvement with dis-              Dryness of vulva                                                                               7 22  19  24  Several                                                                            appearance of dyspareunia.                and marked dys- 14     38  57  Some No withdrawal bleeding.                   pareunia last 2 20     71  64  Few  Discontinued - lack of drug.              years.          20 Biopsy - Late prolif.                                      Biopsy - resting                                                                              endometrium. Moderate                                         endometrium.    mitotic activity.                                               57-A                                                                                     53  0     10   0  Few  Hot flushes gone and feels                Extremely irritable                                                                            7     48  30  Few  much better. Regrets no                   with frequent hot                                                                             14     36  63  Few  more drug. Withdrawal                     flushes.        20     57  75  Few  bleeding moderate - lasted                Biopsy - Intermed.                                                                            20 Biopsy - Late prolif.                                                                          two days. Discontinued-                   prolif. endom.  endometrium. Good   lack of drug.                             Very few mitoses.                                                                             mitotic activity.                                               58-A                                                                                     64  0  74%                                                                               3   0  Many Patient remained in same state.           Palpitation, moder-                                                                            7 16  28  12  Several                                                                            No withdrawal bleeding. Dis-              ate respiratory in-                                                                           14     31  42  Several                                                                            continued - lack of drug.                 sufficiency, easy                                                                             20     57  49  Some                                           fatigability. Widow.                                                                          20 Biopsy - Late prolif.                                      Biopsy - none   endometrium. Slight                                                           mitotic activity.                                             __________________________________________________________________________

                                      TABLE V                                     __________________________________________________________________________    Vaginal Cytology                                                                                                Follicle                                                       Basal     Leuco-                                                                             Stimulating                                   Patient    Age                                                                              Day                                                                              Cells                                                                             CPI                                                                              EI cytes                                                                              Hormone   Remarks                           __________________________________________________________________________      59-A                                                                                     52  0  64%                                                                               6    Several                                                                            0d. 52 m.u.                                                                             Under treatment 50 days,          Hot blushes; nausea                                                                           10 11  27 47 Some 45d.                                                                              6 m.u.                                                                              after 10 days symptoms            in the morning. 20     63 54 Some           disappeared. No prob-             Biopsy - resting                                                                              30     50 62 Some           lems.                             endometrium.    30 Biopsy - Prolif. endom.                                                    Moderate mitotic activity.                                      60-A                                                                                     50  0      7 16 Several                                                                            0d. 112 m.u.                                                                            Under treatment 54 days.          Asthenia and numbness                                                                         10     35 49 Several                                                                            60d.                                                                              12 m.u.                                                                             Marked improvement 10             of hands. Menopause                                                                           20     59 37 Several        to 15 days after start.           4 years ago. Biopsy-                                                                          30     68 49 Some           No problems.                      Early prolif. endom.                                                                          30 Biopsy - Late prolif.                                      Scarce mitotic  endometrium. Moderate                                         activity        mitotic activity.                                               61-A                                                                                     49  0      5  0 Many           Symptoms disappeared after        Hot flushes and slight                                                                        10     27 55 Several        2 weeks. No problems.             irritability. Biopsy-                                                                         20     63 42 Several                                          Resting enodmetrium.                                                                          30     58 60                                                                  30 Biopsy - Late prolif.                                                      endometrium. Slight                                                           mitotic activity.                                               62-A                                                                                     51  0  87%                                                                               3    Many 0d. 64 m.u.                                                                             Symptoms improved 6 or 7          Asthenia and irrita-                                                                          10 2   19 24 Some 30d.                                                                              6 m.u.                                                                              days after start. As-             bility. Biopsy- 20     63 40 Some 60d.                                                                              <6 m.u.                                                                             thenia gone after 15              None.           30     51 66 Some           days. Irritability                                30 Biopsy - Late prolif.    markedly decreased. No                            endometrium. Moderate       problems.                                         mitotic activity.                                               63-A                                                                                     57  0  93%                                                                               0    Many 0d. 120 m.u.                                                                            Hot flushes disappeared           Hot flushes and 10 22  30 12 Several                                                                            30d.                                                                              10 m.u.                                                                             after 7 to 10 days.               itching of genitalia.                                                                         20     36 43 Several        Itching decreased but             Biopsy - resting                                                                              30     42 35 Several        did not vanish. After             endometrium.    30 Biopsy - Intermed. prolif.                                                                             2 weeks tenderness of             endometrium. Slight                                                                           breasts and bloating.                                         mitotic activity.                                                                             She interrupted treat-                                                        ment.                                                           64-A                                                                                     55  0      9 16 Several                                                                            0d. 97 m.u.                                                                             Hot blushes gone in less          Hot blushes and 10     35 55 Several                                                                            30d.                                                                              9 m.u.                                                                              than 7 days. Three days           sporadic diarrhea.                                                                            20     45 38 Several                                                                            60d.                                                                              6 m.u.                                                                              of diarrhea. No                   Biopsy - Early prolif.                                                                        30     56 50 Several        other problems.                   endom. Minimal  30 Biopsy - Late prolif.                                      mitotic activity.                                                                             endometrium. Good mitotic                                                     activity.                                                       65-A                                                                                     63  0  78%                                                                               4    Several                                                                            0d. 185 m.u.                                                                            Hot blushes disappeared,          Hot blushes and 10 32  16 10 Several                                                                            30d.                                                                              <6 m.u.                                                                             and insomnia improved.            insomnia. Biopsy -                                                                            20  3  32 38 Some 60d.                                                                              <6 m.u.                                                                             Feeling very well.                resting endometrium.                                                                          30     29 62 Some                                                             30 Biopsy - Intermed. endom.                                                  Moderate mitotic activity.                                      66-A                                                                                     64  0  86%                                                                               0    Many 0d. 97 m.u.                                                                             Decrease in number of             Moderate irritability                                                                         10 33  30 17 Many 60d.                                                                              12 m.u.                                                                             headaches and intensity.          and headaches, also                                                                           20     33 46 Several        Mild nausea first                 mild diabetes mellitus.                                                                       30  6  45 32 Several        days. Vomited once.               Biopsy - None.  30 Biopsy - Late prolif.                                                      endometrium. Slight                                                           mitotic activity.                                               67-A                                                                                     49  0     10  2 Some 0d. 106 m.u.                                                                            Hot blushes and head-             Hot blushes, nervous-                                                                         10     28 45 Some 7d. 31 m.u.                                                                             aches disappeared                 ness, sporadic head-                                                                          20     62 57 Few  14d.                                                                              25 m.u.                                                                             shortly after start.              aches. Menopause-                                                                             30     58 83 Few  21d.                                                                              13 m.u.                                                                             Nervousness improved              2 years ago. Biopsy-                                                                          30 Biopsy - Late prolif.                                                                        28d.                                                                              6 m.u.                                                                              but not gone.                     Late prolif. endom.                                                                           endometrium. Patchy                                                                             60d.                                                                              <6 m.u.                                 Scarce mitotic  hyperplasia.                                                  activity.                                                                       68-A                                                                                     34  0  98%      Several                                                                            0d. 168 m.u.                                                                            Feeling well. Symptoms            Complete hysterectomy                                                                         10 13  37 52 Some 30d.                                                                              14 m.u.                                                                             gone after 1.0 week.              and ovariectomy.                                                                              20  6  50 43 Some 60d.                                                                              <6 m.u.                                 Had severe      30     67 80 Few                                              hot blushes and 30 Biopsy - None                                              irritability. Biopsy-                                                         not done (hysterectomy)                                                       __________________________________________________________________________

                                      TABLE VI                                    __________________________________________________________________________    Vaginal Cytology                                                                                 Basal     Leuco                                            Patient      Age                                                                              Day                                                                              Cells                                                                             CPI                                                                              EI cytes Remarks                                    __________________________________________________________________________      79-A                                                                                     52  0  42%                                                                               6    Several                                                                             Hot blushes gone after 3 weeks.            Hot blushes and 10 23  14 28 Several                                                                             Remains very nervous.                      nervousness.    20     26 20 Several                                          Biopsy - strophic                                                                             30     46 35 Several                                          endometrium. Cystic                                                                           30 Biopsy - Intermed. prolif.                                 pseudohyperplasia.                                                                            endom. Moderate mitotic                                                       activity.                                                       80-A                                                                                     64  0 93%       Many  Patient feels same and reports             Diarrhea (chronic                                                                             10 37  11    Several                                                                             no change.                                 amoebiasis).    20 30  28    Several                                          Depressed.      30  0  37 28 Several                                          Biopsy - resting                                                                              30 Biopsy - Early prolif.                                     endometrium.    endometrium. Moderate                                                         mitotic activity.                                               81-A                                                                                     50   0    11 31 Several                                                                             Hot blushes gone after 15-20               Marked irritability                                                                           10     35 30 Several                                                                             days. No headaches and moder-              frequent hot    20     63 48 Some  ate decrease in irritability.              flushes, some severe                                                                          30     57 62 Some                                             headaches.      30 Biopsy - Late prolif.                                      Biopsy - resting                                                                              endometrium. Good mitotic                                     endometrium.    activity.                                                       82-A                                                                                     56  0  69%                                                                               0    Many  Patient has not improved.                  Irritability,   10 31  27    Several                                          mild hot blushes and                                                                          20  8  19 Several                                             insomnia - past 2 years.                                                                      30     38 51 Some                                             Diabetic.       30 Biopsy - Late prolif.                                      Biopsy - resting                                                                              endometrium. Patchy                                           endometrium     hyperplasia.                                                    83-A                                                                                     69  0  100%                                                                              0    Many  Good condition. Vaginal in-                Atrophic vaginitis.                                                                           10 71  20    Many  flammation improved.                       Hysterectomy in 20 17  48    Several                                          1953 Marked vari-                                                                             30  4  37    Several                                          cose veins.     30 Biopsy - None                                              Biopsy - hysterectomy                                                           84-A                                                                                     56  0     18  7 Several                                                                             Condition same as before start             Periods of mild 10     36 38 Some  of treatment.                              depression or   20     31 54 Few                                              irritability.   30     40 36 Few                                              Biopsy - Early  30 Biopsy - Late prolif.                                      prolif. endom.  endometrium. Moderate                                         Scarce mitotic  mitotic activity.                                             activity.                                                                       85-A                                                                                     52  0 21  13  6 Several                                                                             Slight improvement of all                  Intense, frequent                                                                             10     39 16 Some  symptoms 10-15 days after                  hot blushes. Occa-                                                                            20     36 44 Some  start of drug.                             sional nausea and                                                                             30     42 29 Some                                             mild headaches. 30 Biopsy - Late prolif.                                      Biopsy - Not    endometrium. Patchy                                           enough material.                                                                              hyperplasia.                                                    86-A                                                                                     53  0     12  3 Several                                                                             Hot blushes disappeared. In-               Insomnia, moderate                                                                            10     36 51 Some  somnia and nervousness un-                 depression, mild                                                                              20     27 50 Some  changed.                                   occasional hot  30     47 26 Some                                             blushes. Biopsy-                                                                              30 Biopsy - Late prolif.                                      Early prolif. endom.                                                                          endometrium. Moderate                                         Scarce mitotic activity.                                                                      mitotic activity.                                               87-A                                                                                     66  0  96%                                                                               0    Many  Vaginitis improved but has not             Kraurosis vulvae,                                                                             10 70  16    Many  disappeared.                               atrophic vaginitis                                                                            20     35 52 Many                                             with moderate   30     28 27 Many                                             leukorrhea.     30 Biopsy - Intermed.                                         Biopsy - mucoid prolif. endom. Slight                                         material.       mitotic activity.                                               88-A                                                                                     49  0     11 21 Some  Hot blushes gone 3 weeks after             Marked frequent hot                                                                           10     30 55 Few   start. Nervousness the same.               blushes and moderate                                                                          20     33 38 Few                                              nervousness.    30     28 47 Few                                              Biopsy - Early prolif.                                                                        30 Biopsy - Late prolif.                                      endom. Slight mitotic                                                                         endometrium. Poor                                             activity        mitotic activity.                                             __________________________________________________________________________

The terms "hot blushes" and "hot flushes" are used synonymously herein.

EXAMPLES XXXI and XXXII

Tables VI and VII below set forth the results of tests in which twogroups of 10 women were fed, respectively, 200 mg./day and 100 mg./dayof F.E.S. In the test summarized in Table VI the drug was administereddaily for 60 days whereas in the test summarized in Table VII the drugwas administered in a regime of 25 days on the drug followed by a 10 dayrest period over a total time period of 180 days. Treatment at 100mg./daily relieved most of the post-menapausal symptoms with theexception of insomnia and depression. Treatment at 200 mg./daily did notrelieve insomnia.

                                      TABLE VII                                   __________________________________________________________________________    Vaginal Cytology                                                                                   Basal      Leuco-                                        Patient        Age                                                                              Day                                                                              Cells                                                                             CPI EI cytes  Remarks                                __________________________________________________________________________      139-A                                                                                      61  0  99%                                                                               0   0 Many Disappearance of symptoms re-            Suffers mild, well-                                                                             10 62  20   7 Many lated to genitalia. Head-                controlled diabetes.                                                                            20 31  27  19 Several                                                                            aches unaffected. No                     Has headache and dry-                                                                           30  7  30  19 Several                                                                            withdrawal bleeding.                     ness with itching of                                                                            60 Late prolif. endom.                                      external genitalia.                                                                             Good mitotic activity.                                        140-A                                                                                      39  0  100                                                                               0   0 Many Patient well. No more hot                Had ovarichystereo-                                                                             10 52  26  13 Some blushes and feels very re-               tomy 8-67. Early  20 21  38  54 Some laxed. No withdrawal                     in October had hot                                                                              30  3  42  80 Few  bleeding.                                blushes and nervousness.                                                        141-A           Early prolif. endom.                                                          Scarce mitotic activity.                                                   56  0      3  11 Several                                                                            Feels better but still has in-           Suffers insomnia and                                                                            10     26  31 Several                                                                            somnia. After second course              slight depression.                                                                              20     38  63 Some had few drops of withdrawal                                30     46  45 Some bleeding.                                                  Late prolif. endom.                                           142-A           Moderate mitotic activity.                                                    Resting endom.                                                             56  0 26   4   0 Several                                                                            No hot blushes since treatment           Marked nervousness                                                                              10 13  16  36 Some started. Nervousness im-                 and occasional hot                                                                              20  2  37  29 Some proved moderately.                       blushes.          30  0  46  38 Some                                                            Intermed. prolif. endom.                                                      Discrete mitotic activity.                                    143-A                                                                                      57    Atrophic endom. with                                                          pseudohyperplasia.                                                          0 17   2   9 Few  No change. No withdrawal                 Mild nervousness. 10     26  33 Few  bleeding.                                                  20     29  57 Few                                                             30     40  58 Few                                                             30 Intermed. prolif. endom.                                                      Moderate mitotic activity.                                 144-A                                                                                      69.   No biopsy.                                               Suffered pyelonephritis                                                                          0 100  0   0 Several                                                                            Patient has mild renal insuffi-          and has atrophy and                                                                             10 64  10   7 Several                                                                            ency. Genitalia wetter with              dryness of vulva. 20 30  29  18 Several                                                                            occasional slight mucorrhea.                               30  7  35  52 Several                                                                            No withdrawal bleeding.                                    60 Early prolif. endom.                                                          Slight mitotic activity.                                   145-A                                                                                            Early prolif. endom.                                                          Discrete mitotic activity.                               Last menstrual period                                                                        49  0     13  26 Some No more hot blushes after one            (LMP) 5/14. Has frequent                                                                        10     35  30 Some week on drug. After end of               hot blushes and feels                                                                           20     48  39 Few  second course had vaginal                nervous. On drug 11/14                                                                          30     66  50 Few  bleeding which was discrete.                               60 Late prolif. endom.                                                           Patchy hyperplasia.                                        146-A                                                                                            Early prolif. endom.                                                          Slight mitotic activity                                  Moderate nervousness                                                                         54  0 14   6  23 Several                                                                            Feels better but has had two             and bouts of diarrhea                                                                           10     21  29 Some episodes of diarrhea. After                                20     38  34 Some second course had moderate               ω           30     35  62 Some withdrawal bleeding for two                                                   days.                                      147-A              Resting endometrium.                                                    51  0 66   3   0 Several                                                                            Hot blushes disappeared shortly          LMP 13 mo. prior to drug.                                                                       10  9  21  38 Some after started drug and irri-             has frequent hot blushes                                                                        20  7  39  32 Some tability improved slightly.              and marked irritability.                                                                        30  1  46  52 Some No withdrawl bleeding.                                     60 Intermed. prolif. endom.                                                      Slight mitotic activity.                                   148-A                                                                                            No biopsy                                                Has mild diabetes                                                                            65  0 100  0   0 Many +                                                                             Besides drug patient received            and atropic vulvular            oandida                                                                            anti-monilial drugs. Vulvo-              vaginitis.                      albicans                                                                           vaginitis cured. No with-                                  10 54   3  17 Many drawal bleeding.                                           20 20  35  23 Several                                                         30 13  56  60 Several                                       __________________________________________________________________________

                                      TABLE VIII                                  __________________________________________________________________________    Vaginal Cytology                                                                                   Basal      Leuco-                                        Patient        Age                                                                              Day                                                                              Cells                                                                             CPI EI cytes  Remarks                                __________________________________________________________________________      119-A                                                                                      62  0  79%                                                                               0   2 Many Itching improved after one week and      Has hypertension and                                                                            10 36   7  10 Several                                                                            disappeared after 3 weeks. No            intense vulvar itching.                                                                         20 17  30  21 Several                                                                            withdrawal bleeding but some             Has diabetes mellitus                                                                           30  6  37  47 Some mucorrhea. Schedule: 25 days             controlled by dieting.                                                                          Endometrial biopsy after 20                                                                      on drug - 10 days off. 180 day           No endometrial biopsy                                                                           days - Intermediate prolif.                                                                      Biopsy - Late Proliferative en-          before treatment. endometrium. Slight                                                                              dometrium. Good mitotic activity                           activity.          At 180 days - Patient feeling                                                 well. Slight withdrawal bleeding                                              for 3 days after four months of                                               treatment.                                 120-A                                                                                      53  0  9  10   3 Some Marked improvement after 5 to 10                                              days                                     Infrequent hot blushes                                                                          10     23  36 Some of treatment. She has been               and moderate nervousness.                                                                       20     51  67 Few  feeling well. Moderate with-             Biopsy - intermed. prolif.                                                                      30     40  58 Few  drawal bleeding after second course      endom. with slight                                                                              20 day Biopsy - Late pro-                                                                        of treatment. Schedule: 25 days          mitotic activity. lif. endom. with moderate                                                                        on drug - 10 days off. At 180                              mitotic activity.  days - Biopsy - Late proliferative                                            endometrium with good mitotic                                                 activity. Feeling well. From                                                  third to sixth cycle no withdrawal                                            bleeding.                                  121-A                                                                                      58  0 14   3  13 Several                                                                            Headache improved but not insomnia.      Has frequent headaches                                                                          10  3  26  17 Some Few drops of blood after third           and insomnia.     20     38  62 Few  course. Schedule: 25 days on             Biopsy - atrophic 30     31  70 Few  drug - 10 days off. Biopsy -             endom. with cystic                                                                              60 day Biopsy - prolif.                                                                          180 days - Proliferative en-             pseudohyperplasia.                                                                              endom. with patchy hyper-                                                                        dometrium with patchy hyperplasia.                         plasia.            180 days. Slight withdrawal                                                   bleeding on fourth and sixth                                                  cycles. Feeling well.                      122-A                                                                                      61  0 64   1  13 Many Feeling a little better. No with-        Moderate irrita-  10 24  27  42 Few  drawal bleeding. Schedule: 25            bility and depression.                                                                          20 18  18  58 Few  days on drug - 10 days off. 180          Biopsy - inactive 30  3  40  37 Few  days biopsy - Late proliferative         endometrium.      30 day Biopsy - prolif.                                                                          endometrium. Feels depressed -                             endom. Slight mitotic                                                                            no withdrawal bleeding.                                    activity.                                                     123-A                                                                                      50  0     16  24 Some After one week of treatment, hot         Frequent intense hot                                                                            10     46  37 Some blushes decreased in number              blushes with abdominal                                                                          20     35  58 Few  and intensity and disappeared            pain and flatulence.                                                                            30     58  65 Few  after 20 days. Flatulence has            Biopsy - early prolif.                                                                          60 day biopsy - Late pro-                                                                        decreased but not disappeared.           endom. with poor  lif. endom. with good                                                                            Schedule: 25 days on drug - 10           mitotic activity. mitotic activity.  days off. 180 day biopsy - Late                                               proliferative endometrium with                                                moderate mitotic activity.                                                    Patient has episodes of abdominal                                             discomfort, otherwise feels well.                                             No withdrawal bleeding.                    124-A                                                                                      68  0 98   0   0 Many Itching improved; vulvar mucosa          Kraurosis vulvae and                                                                            10 80   0   7 Many slightly wet. No withdrawal              moderate itching. 20 53  11  28 Several                                                                            bleeding. Schedule: 25 days on                                                -Biopsy - resting  30 21 35 31 Severa                                         l drug - 10 days off. 180 day            endometrium.      30 day biopsy - Early pro-                                                                       biopsy - Intermediate prolifera-                           lif. endom, with slight                                                                          tive endometrium with poor mitotic                         mitotic activity.  activity. Feeling well. No with-                                              drawal bleeding.                           125A                                                                                       52  0      8   3 Some No headaches and hot blushes dis-        Moderately frequent hot                                                                         10     51  34 Some appeared after one week on               blushes; rare headaches                                                                         20     35  62 Some drug. Minimal withdrawal bleeding        Biopsy - atrophic endom.                                                                        30     60  48 Some after third course. Schedule:            with cystic pseudo-                                                                             30 day biopsy - prolif.                                                                          25 days on drug - 10 days off.           hyperplasia.      endom. with moderate                                                                             180 day biopsy - Late prolifera-                           mitotic activity.  tive endometrium with patchy                                                  hyperplasia. Occasional head-                                                 aches fifth month and slight                                                  withdrawal bleeding on fifth                                                  month.                                     126-A                                                                                      58  0 47   1  13 Many Irritability improved but still de-      Irritability and periods                                                                        10 37   8  20 Several                                                                            pressed over family problems.            of depression.    20 26  17  19 Several                                                                            No withdrawal bleeding. Schedule:        Biopsy - resting en-                                                                            30  8  31  43 Several                                                                            25 days on drug - 10 days off.           dometrium.        30 day biopsy - Early pro-                                                                       180 day biopsy - Late prolifera-                           lif. endom. Moderate mito-                                                                       tive endometrium with discrete                             tic activity.      mitotic activity. Depressed. Has                                              forgotten to take several capsules.                                           No withdrawal bleeding.                    127-A                                                                       55              0 37  8   3  Several                                                                          Remained without significant                                    10 20  17  29 Some change; likely nervousness               Slight nervousness                                                                              20  7  35  23 Some decreased. No withdrawal                 and hypertension. 30 11  47  61 Few  bleeding. Schedule: 25 days              Biopsy - resting  60 day biopsy - late pro-                                                                        on drug - 10 days off. 180               endometrium.      liferative endometrium.                                                                          day biopsy - Early prolifera-                              Moderate mitotic activity.                                                                       tive endometrium with moderate                                                mitotic activity. Irritability                                                -  improved. No withdrawal                                                    bleeding. -                                128-A                                                                                      49  0     14  26 Many Symptoms improved in five days and       Marked and frequent                                                                             10     23  31 Many disappeared after 15 days. Feel-         hot blushes with fre-                                                                           20     28  63 Several                                                                            ing well. Moderate withdrawal            quent moderate headaches.                                                                       30     55  42 Several                                                                            bleeding after second and third          Biopsy - early prolif.                                                                          30 day biopsy - late prolif.                                                                     courses. Schedule: 25 days on            endom. with moderate                                                                            endometrium. Good mitotic                                                                        drug - 10 days off. 180 day              mitotic activity. activity.          biopsy - Proliferative endometrium                                            with patchy hyperplasia. Feeling                                              well. Moderate withdrawal bleeding                                            in fifth and sixth                       __________________________________________________________________________                                         months.                              

EXAMPLE XXXIII

Groups I, II, III and IV each composed of human female patientssuffering postmenapausal syndrome were orally given dosages of 10, 25,50 and 100 mgs./day, respectively, of pure Compound II (LM) having amelting point of about 155° C. for 3 cycles, each cycle consisting oftaking the compound daily for 20 days followed by a 10 day period duringwhich the patient abstains from taking the compound. The results are setforth below.

The pure Compound II (LM) can be separated from a mixture of it and itshigh melting diastereoisomer by solubilizing the mixture in glacialacetic acid and crystallizing out the pure Compound II (LM) inaccordance with the process described in the copending patentapplication of Vernon V. Young, Ser. No. 643,819, filed June 6, 1967,now U.S. Pat. No. 3,574,235 herein incorporated by reference.

    __________________________________________________________________________                                 VAGINAL CYTOLOGY.sup.3                                                                      SUBJECTIVE                                   FSH (units).sup.2  CPI          RESPONSE.sup.4                      GROUP                                                                              DOSE.sup.1                                                                         INITIAL                                                                              90 DAYS TREATMENT                                                                         INITIAL                                                                              20-DAYS                                                                             90 Days                             __________________________________________________________________________    I    10 mg.                                                                             105(144-66)                                                                          25.8(54-6)   5.2(18-0)                                                                           24.7(38-16)                                                                         4 of 10                             II   25 mg.                                                                             102(156-66)                                                                          22.2(36-6)   7.6(19-0)                                                                           40.8(52-26)                                                                         8 of 10                             III  50 mg.                                                                             92.8(138-54)                                                                         12.0(30-6)   5.5(11-0)                                                                           55.2(73-42)                                                                         10 of 10                            IV   100 mg.                                                                             99(150-60)                                                                          11.4(30-6)  11.0(19-0)                                                                           67.5(77-59)                                                                         9 of 10                             __________________________________________________________________________     .sup.1 Dose mg. orally daily for 20 days with a 10 day rest.                  .sup.2 Follicle stimulating hormone assay before and after 3 cycles of        treatment.                                                                    .sup.3 Chorionic pycnotic index.                                              .sup.4 Subjective response as reported to physician by patient.          

It is claimed:
 1. A method of treating cholesterolemia in a human havingcholesterolemia which comprises internally administering to said humanan amount sufficient to alleviate said cholesterolemia of a compoundhaving the formula: ##STR11##
 2. The method of claim 1 wherein saidcompound is administered in an amount of about 0.5 to 20 mg. per day perkilogram of said human body weight.
 3. The method of claim 1 whereinsaid compound is compound II.
 4. The method of claim 3 wherein saidcompound is the diastereoisomer having a melting point of about 178° to180° C.
 5. The method of claim 4 wherein said compound is administeredin an amount of about 0.5 to 20 mg. per day per kilogram of said humanbody weight.